Leaky Expression of Channelrhodopsin-2 (ChR2) In Ai32 Mouse Lines

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dc.contributor.authorPrabhakar, Arthi
dc.contributor.authorVujovic, D.
dc.contributor.authorCui, L.
dc.contributor.authorOlson, W.
dc.contributor.authorLuo, W.
dc.contributor.utdAuthorPrabhakar, Arthi
dc.date.accessioned2019-12-18T22:00:52Z
dc.date.available2019-12-18T22:00:52Z
dc.date.created2019-03-26
dc.description.abstractOptogenetics enables the selective activation of genetically-targeted neuronal populations using light-sensitive ion channels. Genetic strategies using Cre-dependent mouse strains, especially the Ai32 line expressing Channelrhodopsin (ChR2)-EYFP fusion protein, have been a popular means to drive opsin expression in a cell-type specific manner. Here we report a low level of leaky ‘off-target’ (Cre-independent) ChR2-EYFP expression in Ai32/ Ai32 homozygous mice throughout the nervous system. This leaky off-target expression was characterized in multiple prevalent nervous system regions using anti-EYFP immunostaining. Expression of full-length ChR2-EYFP protein was confirmed using immunoprecipitation followed by Western blotting. Notably, light stimulation of these ChR2-EYFP expressing neurons in the spinal cord dorsal horn did not induce detectable photocurrents in juvenile 4-week old mice. Given the wide use of the Ai32 line by many labs, our results suggest researchers should be vigilant of possible off-target ChR2-EYFP expression in their region of interest, especially when generating Ai32/Ai32 homozygotes to drive high levels of ChR2-EYFP expression in adult mice.
dc.description.departmentSchool of Behavioral and Brain Sciences
dc.description.sponsorshipNIH grants (NS083702, NS094224 and NS092297).
dc.identifier.bibliographicCitationPrabhakar, A., D. Vujovic, L. Cui, W. Olson, et al. 2019. "Leaky expression of channelrhodopsin-2 (ChR2) in Ai32 mouse lines." PLOS One 14(3): art. e0213326, doi: 10.1371/journal.pone.0213326
dc.identifier.issn1932-6203
dc.identifier.issue3
dc.identifier.urihttps://hdl.handle.net/10735.1/7132
dc.identifier.volume14
dc.language.isoen
dc.publisherPublic Library of Science
dc.relation.urihttps://dx.doi.org/10.1371/journal.pone.0213326
dc.rightsCC BY 4.0 (Attribution)
dc.rights©2019 The Authors
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.source.journalPLOS One
dc.subjectChannelrhodopsin
dc.subjectChannelrhodopsin 2
dc.subjectAdulthood
dc.subjectAge groups
dc.subjectAnimal tissues
dc.subjectHomozygote
dc.subjectImmunohistochemistry
dc.subjectImmunoprecipitation
dc.subjectMice
dc.subjectNervous system
dc.subjectSpinal Cord Dorsal Horn
dc.subjectWestern immunoblotting
dc.titleLeaky Expression of Channelrhodopsin-2 (ChR2) In Ai32 Mouse Lines
dc.type.genrearticle

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